The Synthesis Of Nitrosyl Ruthenium Complexs With 8-Hydroxy Quinoline Derivatives As Ligands And Their Biological Activity Research

As one of the critical media in mammal,nitric oxide acts as an important role in cell and organ function regulation,the importance of NO in biological systems has caused scientists to pay more attention to research reagents releasing NO in vivo,these agents may release NO under the excitation of heat or light,then NO is delivered to target cells or organs.The metal complexes binding NO have been widespread concerned,not only in coordination chemistry but also in biochemistry and pharmacology.NO can bind to a variety of type of metal ions,thereby forming nitrosyl complexes with different structures,coordination numbers and the electronic properties.Based on these findings,many transition metal nitrosyl complexes as potential carriers of NO have been widely studied,they can release and deliver NO in biological tissue upon irradiation of light.Nitrosyl ruthenium complexes are thought to be potential NO transport agents,due to their thermal stability performance as well as under the condition of light stimulation can release NO.At present,it is a hot scientific research to find effective exogenous NO donors,transport the NO donor to the lesion directionally and release NO in a particular area.In this dissertation,we synthesized three kinds of nitrosyl ruthenium complexes[(CH3)4N][RuCl3(2mqn)(NO)],[(CH3)4N][RuCl3(2cqn)(NO)],[(CH3)4N][RuCl3(qn)(NO)]with the H2mqn,H2cqn and qn as ligands,respectively.The complexes were identified using NMR technology,infrared technology and UV-vis spectroscopy,their physical and chemical properties were also compared;NO radicals release from three kinds of nitrosyl ruthenium complexes upon photoirradiation were quantitatively monitored with NO selective sensor,the preliminary method of quantitative releasing NO was obtained;DNA-EB fluorescence spectrum is studied after the addition of three kinds of nitrosyl ruthenium complexes,their binding mechanism,binding constant and binding-site number were analyzed simply by using the static quenching method,by comparing the binding constants to determine the combined strength of the three kinds of complexes with DNA is:[RuCl3(2cqn)(NO)]->[RuCl3(2mqn)(NO)]->[RuCl3(qn)(NO)]-;the mechanism of photoinduced cleavaging of pBR322DNA with the complexes were investigated,free radical mechanism was analyzed.The interaction of the complexes with HSA,including binding type,binding constant and binding-site number were calculated using fluorescence spectrum,UV absorption spectrum and synchronous fluorescence spectrum;to explore the influence on the activity of cancer cells with the addition of three kinds of complexes.There are mainly seven parts in the following sections:First,the physiological functions of NO,the main NO exogenous donors,the research progress of the photodynamic therapy were introduced and summarized.Second,according to the literature,three kinds of nitrosyl ruthenium complexes[(CH3)4N][RuCl3(2mqn)(NO)],[(CH3)4N][RuCl3(2cqn)(NO)],[(CH3)4N][RuCl3(qn)(NO)]with H2mqn,H2cqn and qn as ligands were synthesized.The location of the characteristic functional groups were measured using infrared spectrum;the chemical shift values of each functional group were measured using NMR technology;the position of characteristic absorption peaks were measured using UV-vis absorption spectrum.Third,the NO release from three kinds of nitrosyl ruthenium complexes under photoirradition were quantitatively monitored by NO selective sensor.With the extension of illumination time,the quantity of released NO increase gradually.Forth,quenching of the fluorescence spectrum of DNA-EB by the complexes were studied by the method of fluorescence spectrum,the model of combination were analyzed,binding constant and binding-site number of interaction of the complexes with DNA were calculated.The combination of the complexes with DNA in binding order[RuCl3(2cqn)(NO)]->[RuCl3(2m qn)(NO)]->[RuCl3(qn)(NO)]-.The cleavage action of the complexes on pBR322DNA under and without photoirradition were studied by agarose gel electrophores,under the condition of photoirradiation,the complexes show obvious cleavage action.Fifth,the combination of the complexes with HSA were analyzed using fluorescence spectrum.Binding constant and binding-site number were obtained by static quenching method.Comparing their binding constant and combining strength of complexes with HSA,the following binding order[RuCl3(2cqn)(NO)]->[RuCl3(2mqn)(NO)]>[RuCl3(qn)(NO)]-was found;use UV-vis spectrum to verify the above conclusion furtherly;the influence of the complexes on the maximum emission wavelength of Tyr and Trp in HSA were analyzed by synchronous fluorescence spectrum,the result suggested that the complexes could affect the conformation of HSA protein.Sixth,the cytotoxic activities of the complexes against MCF-7 and HepG-2 tumors were evaluated under the condition with and without photo irradiation using MTT method,the IC50 values of the complexes on tumor cell were calculated,determine the order of the cytotoxicity of complexes on tumor is:[RuCl3(2cqn)(NO)]->[RuCl3(2mqn)(NO)]->[RuCl3(qn)(NO)]-.The effect of the complexes on HepG-2 cell cycle and cell apoptosis were further analyzed by the method of flow cytometry,the results show that the addition of the complexes will have an impact on tumor cell cycle,can also cause cell apoptosis.